High cell density cultivation of Bacillus subtilis NCIM 2063: Modeling, optimization and a scale-up procedure

Abstract

Bacillus subtilis is a non-pathogenic, sporulating, Gram-positive bacteria with pronounced antimicrobial and metabolic activity and great potential for wide application in various fields. The aim of this paper was to determine the optimum B. subtilis NCIM 2063 growth conditions and to scale up biomass production from shake flasks to a bioreactor level. The critical growth parameters and their interaction effects were studied using Box–Behnken experimental design and response surface methodology. Developed model equations were statistically significant with good prediction capability. It was found that during shake flask cultivation glucose should be added in concentration up to 5 g L-1 in DSM medium, OTR at 10 mol m-3 h-1 and temperature of 33ºC, to achieve the maximum number of viable cells and spores. To scale up the process from shake flasks to the bioreactor level kLa was used as a main criterion. Scale up effect was evaluated by comparing the growth kinetics in the shake flasks and in a laboratory bioreactor. The total number of cells obtained in the bioreactor was 4.57x109 CFU mL-1 which is 1.41 times higher than the number of cells in the shake flasks (3.24x109 CFU mL-1), proving that the scale-up procedure was conducted successfully.

Keywords

• shake flask
• bioreactor
• microbial biomass